Equipment And Reagents For Real-Time PCR

In September 1993 Higuchi et al. published a report in the journal Biotechnology (NY). Titled “Kinetic PCR analysis: real-time monitoring of DNA amplification reactions,” the paper described the technique that would come to be known as Real-Time PCR (for more info, click here). In the original report, the authors took advantage of the DNA binding properties of the chemical ethidium bromide (EtBr) to monitor the accumulation of PCR product in a reaction tube.

During the 1960s a number of groups were actively engaged in studying DNA intercalating agents for use as potential chemotherapy agents. One of the earliest biophysical characterizations of a DNA-intercalator complex was performed in 1965, by Waring, who used UV-visible spectroscopy to analyze the interaction of DNA and EtBr. In 1967, LePecq and Paoletti used fluorescence spectroscopy to study DNA/EtBr interactions. In their work, LePecq and Paoletti demonstrated that a DNA/EtBr complex has a 21-fold greater fluorescence intensity than EtBr that is free in solution. This fact led to the use of EtBr as a stain for DNA in agarose gels, which, eventually, led to its use in Real-Time PCR.

Currently there are a number of different ways to detect PCR product in Real-Time PCR analysis. Primers that have been labeled with fluorogenic tags, as well as new, more sensitive DNA dyes, are now common. In addition, there have been quite a few advances in the machines used to run Real-Time PCR. Take a look below at some of the equipment and reagents currently available for Real-Time PCR analysis.

Featured Products

QuantiTect SYBR Green Kits

QIAGEN

QuantiTect(tm) SYBR® Green PCR and RT-PCR Kits - increase your specificity and sensitivity on any real-time cycler!

The QuantiTect(tm) SYBR® Green PCR Kit (for PCR and two-step RT-PCR) and the QuantiTect SYBR Green RT-PCR Kit (for one-step RT-PCR) enable rapid, accurate, and inexpensive quantification of DNA, cDNA, and RNA targets on all types of real-time cyclers.

HotStarTaq® DNA Polymerase in the master mix of all QuantiTect Kits provides a hot start to the PCR for high specificity, allowing detection of low-copy-number targets. Inaccurate quantification due to SYBR Green I binding to nonspecific products is eliminated.

Real-Time PCR System

Stratagene

Precise temperature control and rapid cycle time for fast results. Four channels with customizable filters. Open platform supports most dyes and chemistries. Optimized for use with Stratagene´s Brilliant® QPCR reagent kits. 96-well format for plates and tubes Fluorogenic Primers

Invitrogen

LUX™ (Light Upon eXtension) Fluorogenic Primers provide a sensitive, specific, and cost-effective detection method for real-time PCR/RT-PCR. Each custom LUX™ Primer Set includes a single-labeled fluorogenic primer and a corresponding unlabeled primer, designed according to the specific sequence of the target gene. This primer set is all you need to perform PCR and real-time detection of the PCR product.

IQ realtime

Bio-Rad

The iCycler iQ real-time PCR detection system is a sophisticated optical system that neatly fits into place directly above the thermal cycler. It covers the widest range of excitation/emission wavelengths available, facilitating the greatest array of fluorescent PCR strategies. When used as part of the iCycler iQ system, the iCycler thermal cycler expands the advantages PCR brings to your research.